Treatment Of Superficial Bladder Cancer Research Articles

Disseminated BCG disease: A case report

Disseminated BCG disease is a rare life-threatening complication of BCG administration, characterised by miliary pulmonary nodules. Presentation can mimic tuberculosis, both in symptoms and on radiological imaging. Here we report a case of disseminated BCG disease following intravesical BCG treatment for superficial bladder cancer that responded to anti-tuberculous treatment and corticosteroids, and briefly review the literature on disseminated BCG disease.

Frequency of Tumor Recurrence: A Strong Predictor of Stage Progression in Initially Diagnosed Nonmuscle Invasive Bladder Cancer

Patients with nonmuscle invasive bladder cancer usually experience multiple instances of tumor recurrence until stage progression occurs. However, it has not yet been fully evaluated whether the timing and/or pattern of tumor recurrence could affect subsequent stage progression. We examined whether the frequency of tumor recurrence provides additional predictive information concerning stage progression. A total of 484 patients with initially diagnosed nonmuscle invasive bladder cancer were identified between 1985 and 2006 at our institution. Median followup was 7.2 years. Frequency of tumor recurrence was analyzed to determine if it affected subsequent stage progression. Of these patients 40 (8.3%) experienced stage progression during followup. Using Kaplan-Meier analysis subsequent stage progression could be most strongly predicted in patients with a recurrence rate of 1 or more per year during the first 2 years, although similar results were observed for various cutoff periods and recurrence rates. The 10-year progression-free survival rate was 58.0% in patients with a recurrence rate of 1 or more per year and 93.3% in their counterparts (p <0.001). Multivariate analysis demonstrated that the appearance of tumor grade 3 (p = 0.027, risk ratio 2.36), carcinoma in situ (p = 0.045, risk ratio 2.44) and a recurrence rate of 1 or more per year during the first 2 years (p <0.001, risk ratio 7.40) were independent risk factors for subsequent stage progression. Frequency of tumor recurrence is a strong predictor of subsequent stage progression in patients initially diagnosed with nonmuscle invasive bladder cancer. More appropriate followup and aggressive treatment due to a higher malignant potential for stage progression might be recommended in patients with a recurrence rate of 1 or more per year during the first 2 years.

Intravesical gemcitabine for treatment of superficial bladder cancer not responding to Bacillus Calmette-Guérin vaccine

Intravesical Bacillus Calmette-Guerin (BCG) vaccine is the mainstay of treatment and prophylaxis in superficial bladder cancer (SBC) as it reduces tumor recurrence and disease progression. About one-third of patients do not respond to BCG. The aim of this study was to determine the efficacy of intravesical gemcitabine in patients with BCG-refractory SBC. Twenty three patients with SBC; TaG3, T1G2-G3 or carcinoma in situ (CIS), refractory (after at least 2 courses of intravesical BCG) or intolerant to intravesical BCG therapy were included. Two weeks after complete tumor resection, patients received intravesical gemcitabine twice weekly at a dose of 2.000 mg/100 ml normal saline for 6 consecutive weeks. Two months after the last dose, recurrence-free patients underwent cystoscopy, urinary cytology and 6 random bladder biopsies. Thereafter, patients were evaluated by the same measures every 3 months, as long as there was no recurrence. Patients with complete response (negative cytology and random biopsies) at the first follow-up cystoscopy received a similar maintenance dose once weekly for another 6 weeks. Twenty one patients completed the study: 15 males and 6 females with a mean age of 48.1 (38–72) years. The follow-up was 15 months (range 2–19 months). Thirteen (61.9%) patients were recurrence-free after a mean of 17 months. Superficial recurrences were detected in 6 (28.6%) patients and progression by stage in 2 patients (9.5%). During follow-up, 8 patients had tumor recurrences and 2 had progression to a higher stage. The median recurrence-free time was 14.7 months (5–19 months). The drug was well tolerated and side-effects were mild in all patients, except two: one had easily controlled hematuria and the other had leucopenia. In properly selected patients, gemcitabine seems to be a promising option in the management of high-risk BCG-refractory SBC, especially in those who refuse or are unfit for cystectomy. Long-term efficacy and the role of maintenance therapy have to be properly studied.

Granulomatous Hepatitis after BCG Intravesical Immunotherapy

Purpose: An 80 y/o male presented with a progressively worsening fever, chills and night sweats 10 days after he received his fifth intravesical BCG instillation for his low-grade bladder carcinoma. Review of systems was remarkable only for mild dysuria and hematuria. Physical examination revealed a temperature of 102°F. He had no hepatosplenomegaly or lymphadenopathy. Labs were significant for leukopenia (3000/μL) and elevated LFTs (alkaline phosphatase [ALP]-378, AST-70, ALT-26 U/L). Prior to presentation, Isoniazid was initiated by his urologist for presumptive BCG infection. Rifampin and levofloxacin were added upon admission to the hospital. Routine blood and urine cultures did not yield any pathogens and serology for viral, bacterial, and fungal infections were negative. Chest x-ray was unremarkable but a chest-CT showed diffuse nodular infiltrates suspicious for Mycobacterium infection. An USG-guided liver biopsy was performed for a persistently elevated ALP, and revealed granulomatous hepatitis with non-caseating epitheloid granulomas and lymphocytic cholangitis. Gram, AFB, GMS, FITE and immunohistochemical stains did not detect any organisms, and tissue culture was negative. Worsening pancytopenia prompted a bone marrow biopsy that revealed several large granulomas. The patient gradually improved, and his fever subsided within several days of anti-tuberculous therapy. Liver abnormalities resolved during the following week. He completed a 6-month course of isoniazid and rifampin, and had no complications at 1-year follow-up. Intravesical BCG is an effective treatment for superficial bladder cancer (success rate: 63-100%). Minor complications including hematuria, cystitis, fever (<38.5°C) and malaise are quite common, whereas major complications such as both local (granulomatous prostatitis, epididymo-orchitis) and systemic reactions (fever >39.5°C, sepsis, cytopenias, granulomatous hepatitis and pneumonitis) occur occasionally. Granulomatous hepatitis is a rare (<0.4%) and serious complication after BCG instillation, with unclear pathogenesis. It has been considered a hypersensitivity reaction to BCG, but cases have been reported in which mycobacteria were seen on staining and mycobacterial DNA was detected in liver tissue suggesting a liver infection after haematogenous dissemination of BCG, rather than hypersensitivity. There have been no prospective studies to evaluate the optimal treatment for BCG infection. In severe systemic cases, some data supports the administration of three anti-tuberculous drugs including isoniazid, rifampin and ethambutol for at least 6 months. The addition of corticosteroids during initial therapy may assist in a faster resolution of inflammatory complications.

Mycobacterium bovis BCG: the importance of an accurate identification in the diagnostic routine

M. bovis BCG is used clinically in the immunotherapy treatment of superficial bladder cancer to prevent progression to invasive disease, leading in some cases to a severe localized inflammation or disseminated infections. For this reason, an accurate and early identification of this particular microorganism is clinically relevant.We describe a case-report of bladder cancer with a urine culture-positive for mycobacteria initially diagnosed as MTB complex infection and later identified as BCG disease by molecular methods.

Conditions causing gemcitabine crystallization

Intravesical delivery of chemotherapy agents is used very commonly for the treatment of superficial bladder cancer. We recently completed a phase II study of intravesical gemcitabine in which an alkaline adjusted gemcitabine preparation was used initially, based on very early phase I studies. However, crystallization was noted in some of the pre-prepared syringes, which prompted us to investigate the conditions under which gemcitabine crystallized. Gemcitabine was prepared in syringes in triplicate and conditions were varied with respect to pH, temperature, and duration. Samples were observed for up to 48 h for the development of crystallization. High-performance liquid chromatography analysis of gemcitabine concentrations was undertaken for all samples. Crystallization of gemcitabine was favored under conditions of bicarbonate treatment and lowering of temperature. However, the process was reversible, as demonstrated by recovery of gemcitabine concentrations in samples brought back to room temperature. Crystallization resulted in reduction of gemcitabine concentrations in the pre-prepared syringes. Gemcitabine solutions may be associated with crystallization if the native pH is increased with the addition of sodium bicarbonate, and samples are stored in a cold environment.

Safety and Efficacy of Intravesical Bacillus Calmette-Guérin Plus Interferon α-2b Therapy for Nonmuscle Invasive Bladder Cancer in Patients With Prosthetic Devices

Patients with bladder cancer who have prosthetic devices, such as a cardiac pacemaker, artificial heart valve or orthopedic hardware, and who undergo intravesical bacillus Calmette-Guérin therapy are theoretically at higher risk for complications, including bacterial seeding of pacemaker wires or orthopedic hardware, and at further risk for infective endocarditis. We assessed the safety and efficacy of bacillus Calmette-Guérin plus interferon α-2b therapy in patients with nonmuscle invasive bladder cancer and a pacemaker, artificial heart valve or orthopedic hardware. We evaluated 1,045 patients with nonmuscle invasive bladder cancer enrolled in a multicenter American phase II trial of bacillus Calmette-Guérin plus interferon α-2b therapy, including 143 with a prosthetic device (pacemaker in 87, artificial heart valve in 13 and orthopedic hardware in 43). Weekly physician toxicity assessments and standard adverse effect reporting were done. No patient had infective endocarditis or hardware infection. One patient with a pacemaker, 2 with orthopedic hardware and none with an artificial heart valve required treatment cessation for fever greater than 102.5F. All defervesced within 24 hours and had no long-term sequelae. Due to intolerable, nonlife threatening side effects 12 patients with a pacemaker, 2 with orthopedic hardware and 1 with an artificial heart valve stopped treatment. Of the remaining patients with a prosthesis 99 and 24 stopped treatment due to intolerable, nonlife threatening and serious side effects, respectively. Patients with a pacemaker, artificial heart valve or orthopedic hardware were no more likely than the general population to have infection or fever, or discontinue treatment due to side effects. These patients should not be excluded from intravesical bacillus Calmette-Guérin plus interferon α-2b therapy for nonmuscle invasive bladder cancer.

Effect of mitomycin C on concentrations of vascular endothelial growth factor and its receptors in bladder cancer cells and in bladders of rats intravesically instilled with mitomycin C

• To examine, using in vitro and in vivo models, the largely unexamined effect of mitomycin C (MMC), an effective intravesical treatment for superficial bladder cancer and carcinoma in situ, on expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR-2), which mediates many of the angiogenic properties of VEGF. • To measure, as a positive control, concentrations of the inhibitor of apoptosis, survivin, as an assessment of MMC effectiveness. • To measure MMC-induced changes in proliferation in the presence and absence of VEGF-A small interfering RNA (siRNA). • After treatment with increasing MMC concentrations (5-200 µg/mL), we measured proliferation, as well as VEGF, survivin, VEGF receptor-1 (VEGFR-1) and VEGFR-2 concentrations in RT-4 and T-24 bladder cancer cells. • The effect of pre-treatment of VEGF siRNA and survivin siRNA on MMC-induced decreases in proliferation was measured. • Urinary VEGF concentrations and bladder and kidney concentrations of VEGF-A, VEGFR-1, VEGFR-2 and interleukin-6 (IL-6) mRNA were measured in rats intravesically instilled with saline or MMC (200 µg/mL). • Although MMC treatment inhibited cell proliferation and decreased survivin mRNA expression in T-24 and RT4 cells, MMC (12-50 µg/mL) increased VEGF-A mRNA and VEGFR-2 mRNA and protein expression. • Pre-treatment with VEGF-A siRNA or survivin siRNA before MMC treatment reduced proliferation more than MMC alone. • MMC-induced reductions in proliferation were reduced additively by pre-treatment with survivin siRNA, but were potentiated by pre-treatment with VEGF-A siRNA. • VEGFR-2 mRNA and protein concentrations and urinary VEGF concentrations were increased in bladders of rats instilled with MMC. • Intravesically instilled MMC increases urinary VEGF and bladder VEGFR-2 protein and mRNA in rats. • MMC increases VEGF mRNA and VEGFR-2 protein and mRNA concentrations in bladder cancer cells. Therefore, we speculate that MMC could increase the angiogenic potential of both cancer and normal cells. • In cancer cells this effect is largest at lower MMC concentrations. • Combining MMC with agents that reduce EGF concentrations could be of value in treatment of transitional cell carcinoma of the bladder (TCC).

Abstract 5614: Interleukin-10 inhibits the induction of macrophage cytotoxicity against bladder cancer cells by Mycobacterium bovis bacillus Calmette-Guérin (BCG)

Abstract Intravesical BCG therapy has been used for the treatment of superficial bladder cancer for three decades. However, the mechanisms underlying BCG therapy currently remain elusive. We have focused on BCG induction of macrophage cytotoxicity against bladder cancer cells. Previously, we observed that macrophages are cytotoxic to bladder cancer cells upon stimulation with BCG in vitro. However, we found that macrophages from C57BL/6 mice are less potent than those from C3H/HeN mice for the killing of both bladder cancer MBT-2 cells and MB49 cells. This study was to determine whether macrophage-derived interleukin (IL)-10 is responsible for this discrepancy between these two commonly used murine bladder cancer models. Thioglycollate-elicited peritoneal exudate cells were harvested from mice 3 days after intraperitoneal injection of 2 ml 3% thioglycollate. Cells were incubated for 3 hours and non-adherent cells were removed after incubation. The remaining cells were 96% F4/80-positive cells as determined by flow cytometry. The prepared macrophages were then used for the analysis of BCG induction of cytotoxicity, cytokines, and nitric oxide (NO) in vitro. Studies demonstrated that BCG-stimulated C57BL/6 macrophages produced a substantially high level of IL-10 compared to BCG-stimulated C3H/HeN macrophages. This high IL-10 production correlated with reduced production of tumor necrosis factor (TNF)-α, IL-6 and NO in BCG-stimulated C57BL/6 macrophages. Neutralizing endogenous IL-10 during BCG stimulation increased C57BL/6 macrophage cytotoxicity against MB49 cells by 3.2-fold, along with increased production of TNF-α by 6.4-fold and NO by 3.6-fold, respectively. Macrophages from genetically modified C57BL/6 mice lacking IL-10 (IL-10−/−) also exhibited increased killing of MB49 cells and production of TNF-α and NO upon BCG stimulation. In addition, supplementation of exogenous recombinant IL-10 reduced BCG-induced C3H/HeN macrophage cytotoxicity against both MBT-2 cells and MB49 cells in a dose-dependent manner. Taken together, our results reveal the inhibitory role of IL-10 in BCG-induced macrophage cytotoxicity, suggesting that blockage of IL-10 may potentially enhance the effect of BCG in the treatment of bladder cancer patients, particularly for BCG non-responders who are often associated with high IL-10 production in response to BCG treatment. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5614.

The Effects of Intravesical Chemoimmunotherapy with Gemcitabine and Bacillus Calmette-Guérin in Superficial Bladder Cancer: A Preliminary Study

This prospective study investigated the long-term effects of intravesical chemoimmunotherapy with gemcitabine (GEM) and bacillus Calmette-Guérin (BCG; n = 36) versus BCG alone (n = 51) for the treatment of superficial bladder cancer. For the chemoimmunotherapy (GEM + BCG) group, GEM (1000 mg) was instilled immediately after transurethral resection of bladder tumour (TURBT) and again (2000 mg) 1 week later. From 2 to 7 weeks after TURBT, BCG was instilled into the bladder of all patients once weekly. The recurrence-free period of the GEM + BCG group (24.13 months) was significantly longer than that of the BCG monotherapy group (19.81 months). The overall recurrence rate was similar between the groups, although at 6 and 9 months post-TURBT, GEM + BCG produced a significantly lower rate of recurrence compared with BCG alone. This study suggests that intravesical chemoimmunotherapy with GEM + BCG is effective in reducing early tumour recurrence and in prolonging the recurrence-free period of superficial bladder cancer.

Should Patients With Primary Upper Urinary Tract Cancer Receive Prophylactic Intravesical Chemotherapy After Nephroureterectomy?

We assessed the efficacy of prophylactic intravesical chemotherapy for primary upper urinary tract urothelial cancer after nephroureterectomy during long-term followup. From January 1985 to June 2007, 196 patients with primary upper urinary tract urothelial cancer were included in this study. Patients were divided into group 1-31 who received intravesical epirubicin instillation, group 2-27 who received intravesical mitomycin C instillation and group 3-138 who did not receive prophylactic instillation after nephroureterectomy. We compared the bladder tumor recurrence rate, number of recurrence episodes, time to first bladder tumor recurrence, tumor type, percent of patients with cystectomy and percent who died of urothelial cancer, and the recurrence-free survival rate. Of the 196 patients 73 had subsequent bladder recurrence at a mean followup of 55.6 months. There were no significant differences in recurrence type, mean number of bladder tumor recurrences, percent of patients with cystectomy and the cancer specific survival rate. The bladder recurrence rate was lower in group 1 and 2 than in group 3. Mean time to first bladder tumor recurrence was longer in groups 1 and 2. Kaplan-Meier curves of recurrence-free survival rates were significantly increased in groups 1 and 2. Intravesical instillation of epirubicin or mitomycin C appears to be well tolerated and effective for preventing bladder recurrence and prolonging time to first bladder recurrence. Patients should receive prophylactic intravesical chemotherapy after nephroureterectomy.

ACUTE RESPIRATORY DISTRESS SYNDROME SECONDARY TO DISSEMINATED MYCOBACTERIUM BOVIS FROM THERAPEUTIC INTRAVESICULAR INSTILLATION OF BACILLUS CALMETTE-GUERIN VACCINE

INTRODUCTION: Intravesicular Bacillus Calmette-Guerin (BCG) treatment is a common treatment for superficial bladder cancer and is typically not associated with significant systemic side effects. Rarely, systemic dissemination and sepsis have been reported. Here we present a fatal case of dissemination following treatment with intravesicular BCG.

Incidental Detection of Granulomatous Prostatitis by F-18 FDG PET/CT in a Patient with Bladder Cancer

A 62-year-old man with transitional-cell carcinoma of the bladder had undergone transurethral resection and adjuvant treatment with several intravesical instillations of bacillus Calmette-Guerin 10 months earlier. The patient was referred for F-18 FDG PET/CT after detection of mediastinal lymphadenopathy on follow-up CT. Beside moderately increased FDG uptake in bilateral hilar and bronchopulmonary lymph nodes, focally intense FDG uptake was detected in the right lobe of the prostate. Histopathologic evaluation revealed granulomatous prostatitis which is a recognized rare complication of intravesical instillation of BCG during the treatment of superficial bladder cancer.

Intravesical immunotherapy of superficial bladder cancer with chitosan/interleukin-12.

Intravesical BCG has been used successfully to treat superficial bladder cancer for three decades. However, 20% to 30% of patients will fail initial BCG therapy and 30% to 50% of patients will develop recurrent tumors within 5 years. Alternative or complementary strategies for the management of superficial bladder cancer are needed. Interleukin-12 (IL-12) is a potent T(H)1 cytokine with robust antitumor activity and the ability to potentiate immunologic memory. Unfortunately, intravesical IL-12 did not show antitumor efficacy in a recent clinical study of patients with recurrent superficial bladder cancer. We hypothesized that coformulation of IL-12 with chitosan, a biocompatible, mucoadhesive polysaccharide, could improve intravesical IL-12 delivery and provide an effective and durable alternative for the treatment of superficial bladder cancer. In antitumor studies, 88% to 100% of mice bearing orthotopic bladder tumors were cured after four intravesical treatments with chitosan/IL-12. In contrast, only 38% to 60% of mice treated with IL-12 alone and 0% treated with BCG were cured. Antitumor responses following chitosan/IL-12 treatments were durable and provided complete protection from intravesical tumor rechallenge. Urinary cytokine analysis showed that chitosan/IL-12 induced multiple T(H)1 cytokines at levels significantly higher than either IL-12 alone or BCG. Immunohistochemistry revealed moderate to intense tumor infiltration by T cells and macrophages following chitosan/IL-12 treatments. Bladder submucosa from cured mice contained residual populations of immune cells that returned to baseline levels after several months. Intravesical chitosan/IL-12 is a well-tolerated, effective immunotherapy that deserves further consideration for testing in humans for the management of superficial bladder cancer.

PDT combined with Intravesical BCG instillation would form an autovaccine for bladder cancer?

Bladder cancer is the second most common urologic malignancy after prostate cancer. Intravesical BCG as a treatment of superficial bladder cancer (SBC) has been used by urologists for over 30 years. Recently, photodynamic therapy (PDT), which uses a red laser and a photosensitive drug to destroy cancer cells, has been showed encouraging results in SBC treatment. However, BCG and PDT are applied to treatment of SBC alone. Currently, cancer vaccines are made in vitro. Several studies confirmed that tumour cells treated in vitro by PDT can be used for generating potent cancer vaccines, which were more effective than other modes of creating whole tumor vaccines, i.e., UV or ionizing irradiation [Gollnick SO, Vaughan L, Henderson BW. Generation of effective antitumor vaccines using photodynamic therapy. Cancer Res 2002;62:1604-8]. Moreover, BCG is a pleiotropic immune stimulator oriented to cellular immunity. We thought that: after PDT destroyed targeted tumor cells on a large scale, Intravesical BCG could elicit and amplify the immune responses, which would directly form an in situ autovaccine in vivo against the primary tumor and metastases at distant sites. In this paper, we propose that the combination of Intravesical BCG and PDT would be a promising new modality for bladder cancer.

A phase I study of intravesical Ad-IFN in superficial bladder cancer patients

e16048 Background: Intravesical BCG (Bacillus Calmette-Guérin) is the gold standard for treatment of superficial bladder cancer. Recurrence is a significant problem. Intravesical interferon α (IFN) demonstrated complete response (CR) in ∼40% patients with most relapsing &lt;1 year. Intravesical Ad-IFN takes advantage of the bladder to allow prolonged exposure to high concentrations of IFN; this is expected to potentiate durable therapeutic responses. Syn3 is an excipient used to increase adenoviral vector (Ad) transfection. Methods: The phase I study is a nonrandomized, open-label, rising-dose, multi-center study of Ad-IFN in patients with papillary bladder cancer, or carcinoma in situ refractory to BCG. Patients with T1 were not enrolled unless they refused cystectomy. A minimum of 3 subjects were enrolled at each dose level. Adverse events and lab abnormalities were graded according to CTC version 3 criteria. The occurrence of treatment-related Grade 3 or Grade 4 toxicity is considered dose-limiting toxicity (DLT). The maximum tolerated dose is defined as the dose which results in DLT in ≤1/6 subjects. Urine from pre-treatment, posttreatment days 0–7, 10, 14, and 28 were tested for IFN, Syn3, Ad-IFN DNA, and research biomarkers. Blood was assayed for IFN, Syn3, Ad-IFN DNA, anti- adenovirus and anti-IFN antibodies. Results: Intravesical administration of 3×109, 1×1010, 3x1010, 1×1011, or 3×1011 particles/ml of Ad-IFN (75ml) in 1mg/ml Syn3 is safe with only minor initial urgency controlled with anticholinergic medication. Of the 13 evaluable patients, 6 are CR, defined as a negative cytology and biopsy at 3 months. CR patients were offered a second dose. Response duration is variable with longest &gt;1 year. Urinary IFN was seen for all patients in a dose-dependent fashion. TRAIL, IP-10, M65, and M30 were also seen. Conclusions: Initial safety and response activity justify further clinical development. [Table: see text]

Recombinant bacillus Calmette-Guérin (BCG) expressing interferon-alpha 2B enhances human mononuclear cell cytotoxicity against bladder cancer cell lines in vitro.

The proper induction of cellular immunity is required for effective bacillus Calmette-Guérin (BCG) immunotherapy of bladder cancer. It has been known that BCG stimulation of human peripheral blood mononuclear cells (PBMC) leads to the generation of effector cells cytotoxic to bladder cancer cells in vitro. To improve BCG therapy, we previously developed human interferon (IFN)-alpha 2B secreting recombinant (r) BCG (rBCG-IFN-alpha). We demonstrated that rBCG-IFN-alpha augmented T helper type 1 (Th1) cytokine IFN-gamma production by PBMC. In this study, we further investigated whether rBCG-IFN-alpha could also enhance PBMC cytotoxicity toward bladder cancer cells. PBMC were prepared from healthy individuals, left alone or stimulated with rBCG-IFN-alpha or control MV261 BCG, and used as effector cells in (51)Cr-release assays. Human bladder cancer cell lines T24, J82, 5637, TCCSUP, and UMUC-3 were used as target cells. To determine the role of secreted rIFN-alpha as well as endogenously expressed IFN-gamma and IL-2 in inducing the cytotoxicity, PBMC were stimulated with rBCG-IFN-alpha in the presence of neutralizing antibodies to IFN-alpha, IFN-gamma or IL-2. To determine the role of natural killer (NK) and CD8(+) T cells in inducing the cytotoxicity, both cell types were isolated after BCG stimulation of PBMC and used as effector cells in (51)Cr-release assays. Non-stimulated PBMC showed basal levels of cytotoxicity against all target cell lines tested. MV261 BCG increased the PBMC cytotoxicity by 1.8- to 4.2-fold. rBCG-IFN-alpha further increased the PBMC cytotoxicity by up to 2-fold. Elevated production of IFN-gamma and IL-2 by PBMC was observed after rBCG-IFN-alpha stimulation. Blockage of IFN-alpha, IFN-gamma or IL-2 by neutralizing antibodies during rBCG-IFN-alpha stimulation reduced or abolished the induction of PBMC cytotoxicity. Both NK and CD8(+) T cells were found to be responsible for the enhanced PBMC cytotoxicity induced by rBCG-IFN-alpha with the former cell type being more predominant. rBCG-IFN-alpha is an improved BCG agent that induces enhanced PBMC cytotoxicity against bladder cancer cells in vitro. This rBCG strain may serve as an alternative to BCG for the treatment of superficial bladder cancer.

MP-2.04: Broadhesive Coated Nanoparticles Loaded with Mitomycin C for the Effective Chemotherapy of Superficial Bladder Cancer

To overcome difficulties in the establishment of a suitable and effective intravesical chemotherapeutic drug concentration due to the periodical discharge of bladder for the treatment of superficial bladder cancer.

Immunomodulatory effects of recombinant BCG expressing pertussis toxin on TNF-alpha and IL-10 in a bladder cancer model

BackgroundSince successful treatment of superficial bladder cancer with BCG requires proper induction of Th1 immunity, we have developed a rBCG-S1PT strain that induced a stronger cellular immune response than BCG. This preclinical study was designed to compare the modulatory effects of BCG and rBCG-S1PT on bladder TNF-α and IL-10 expression and to evaluate antitumour activity.MethodsFor Experiment I, the MB49 bladder cancer cell line was used in C57BL/6 mice. Chemical cauterization of the bladder was performed to promote intravesical tumor implantation. Mice were treated by intravesical instillation with BCG, rBCG-S1PT or PBS once a week for four weeks. After 35 days the bladders were removed and weighed. TNF-〈 and IL-10 cytokine responses were measured by qPCR. Experiment II was performed in the same manner as Experiment I, except the animals were not challenged with MB49 tumor cells. Results: rBCG-S1PT immunotherapy resulted in bladder weight reduction, compared to the BCG and control group. There were increases in TNF-α in the BCG-treated group, as well as increases in TNF-α and IL-10 mRNA in the rBCG-S1PT group.ConclusionThese data indicate a significant reduction of bladder tumor volume for the rBCG group, compared to the BCG and PBS groups. This suggests that rBCG could be a useful substitute for wild-type BCG and that the potential modulation between TNF-α and IL-10 cytokine productions may have therapeutic value.

Efficacy of bacillus Calmette‐Guérin in the treatment of superficial bladder cancer: The impact of previous intravesical treatment

To clinically evaluate the role of intravesical bacillus Calmette-Guérin (BCG) in the management of superficial bladder cancer, focusing on dose, age, high grade/stage, and pretreatment episodes with other therapies in long-term follow-up patients. A total of 213 patients with superficial (Ta-T1) transitional cell carcinoma of the bladder received 6-8 weekly instillations of 40, 60 or 80 mg of Tokyo strain BCG. Further 6-8 monthly applications were given in some cases. Recurrence-free and progression-free survival rates were compared with reference to dose, age, previous treatment, grade, stage, and high risk categories. Overall recurrence-free rates for 3, 5, and 10 years were 70.8%, 67.1%, and 57.6%, with progression-free survival rates of 94.5%, 93.6%, 90.6%, respectively. In univariate analyses, younger cases demonstrated higher recurrence-free survival rates, but without statistical significance (P = 0.1229). Recurrence-free rates were significantly higher for cases without previous intravesical treatment (P = 0.0010). There was no significant BCG dose-dependence. High grade and high stage patients were confirmed as having higher recurrence and progression rates. Intravesical instillation of BCG is the most effective in the long term when chosen as the initial prophylactic therapy for the prevention of recurrence in superficial bladder cancer.